Cell line genetic modification is rapidly advancing due to the establishment of genome editing technology. Using this technology, it’s now possible to disrupt or insert specific genes. Our company also offers a genetic modification service for cell lines using genome editing.

Our successful experiences
HEK293/A549/U937/YTN16/MDCK/CMT167/MOC2/Jurkat/C1498/THP-1/L929

How to edit cellular genome by CRISPR-Cas9 system

1. Preparation of cell lines (1 week)
   Upon receiving the requested cell line, we will expand the cells from the frozen vial and use them for genetic modification with the CRISPR/Cas9 system. The provided cell line will be tested for mycoplasma before use.
2. Genome editing
   Using CRISPR, we perform genome editing on the prepared cell lines. Please note that we have obtained a license from the Broad Institute (Cambridge, MA, USA) for the use of CRISPR.
   • gRNA Preparation (1-2 months)
     We will select highly specific gRNA sequences that are common to the sequences found using both “Optimized CRISPR Design” (http://crispr.mit.edu/) and “CRISPRdirect” (http://crispr.dbcls.jp/). To confirm the sequence information, we will also perform sequence analysis of the gRNA-targeted region.
   • gRNA Preparation and RNP Delivery (1 month)
     We will prepare an RNP complex consisting of the gRNA and Cas protein targeting the gene of interest and introduce it using a 4D-NucleofectorTM (LONZA).
     After RNP delivery, we will perform genotype analysis (PCR screening) on each well of the cell line to confirm whether genetic modification has occurred.